FIGURE 2.

Casodex disrupts telomeric complexes without affecting the expression of telomeric proteins. A, LNCaP cells treated with 100 μm Casodex for 0, 3, 10, 24, or 48 h were immunostained for 53BP1. 53BP1 foci were counted, and cells were categorized as having ≤5 or >5 foci/cell. ∼200 cells in each treatment group were scored in three independent experiments. B, total RNA from LNCaP cells treated with Casodex was extracted using TRIzol (Invitrogen), and RT-PCR was performed as described previously (32) to measure mRNA levels. Sequence-specific primers for TIN2, TRF1, TRF2, TPP1, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as described previously (11), and for AR, 5-tcagttcacttttgacctgctaa-3′ (forward) and 5′-gtggaaatagatgggcttga-3′ (reverse) primers were used. C, whole cell lysates of LNCaP cells treated with Casodex were subjected to Western blot analysis to determine TIN2, TRF1, TRF2, AR, and glyceraldehyde-3-phosphate dehydrogenase (loading control) protein levels.